文档介绍：survivin启动子的克隆及其在前列腺癌细胞系中的活性鉴定
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作者:骆晓梅,张南征,刘家云,苏明权,郝晓柯
【摘要】目的克隆survivin启动子片段,并检测其在人前列腺癌细胞中的转录活性,为该启动子在前列腺癌靶向基因治疗中的应用提供实验依据。方法 PCR方法扩增survivin基因启动子S1pro和S2pro,克隆入pGL3-Basic,分别构建重组质粒pGL3-S1pro和pGL3-S2pro,脂质体转染前列腺癌细胞和Chang liver肝细胞,检测survivin启动子在细胞中的转录活性。结果 survivin基因启动子在前列腺癌细胞中均具有较强活性,其中S2pro活性明显高于S1pro,达到CMV启动子活性的三分之一。结论 survivin启动子在前列腺癌细胞中具有较强启动活性,有可能成为新的前列腺癌靶向性基因治疗工具。
【关键词】前列腺癌;基因;survivin;启动子;基因治疗
Abstract: Objective To clone DNA sequence of survivin promoter, and detect its transcriptional activities in human prostate cancer cells. Methods The fragment of survivin promoter was acquired by PCR amplification and was inserted into pGL3-Basic to reconstruct binant plasmids pGL3-S1pro and pGL3-S2pro, respectively. The reconstructed plasmids were transiently transfected into human prostate cancer cell lines LNCap, PC-3 and normal Chang liver cells. The transcriptional activities of survivin promoter in various cel1s were determined by detection of the expression of luciferase. Results Survivin promoter had transcriptional activities in all prostate cancer cell lines and the transcriptional activity of the S2pro was much higher than the S1pro and reached a level of one third of the transcriptional activity of the CMV promoter. Conclusion The survivin promoter cloned in this study was more highly activated in prostate cancer ce